Micro propagation, callus induction and root culture of hyoscyanzus niger l., a temperate medicinal plant
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Date
2003-05
Authors
Ee May, Lai
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Abstract
A two step sterilisation method was found to be the best way to obtain the
highest percentage of aseptic in viiro seedlings of Hyoscyamus niger. This method
involved sterilising the seeds with 10% Clorox® for 15 minutes m the first stage
followed by 5% Clorox® for 10 minutes in the second stage. By using this method,
71.3% of aseptic seeds germinated and all the in vitro seedlings (100%) survived.
Multiple shoots were successfully induced from the apical shoots using MS medium
supplemented with 8.0 mg!L BA and 0.5 mg!L ffiA. The separated shoots produced
roots on basic MS medium to form complete plantlets. The leaf explants were found to
be the best tissue for the induction of callus. The leaf explants cultured on MS medium
supplemented with 6.0 mg!L picloram produced the most callus with soft and friable
type. The compact type of callus was induced from the leaf explants that were cultured
on MS plus 2.0 mg!L picloram. The compact callus showed the presence of both
hyoscyamine and scopolamine alkaloids. Roots were successfully induced on solid MS
medium supplemented with 2.0 mg/L IBA. In liquid medium, the most suitable
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medium for root pr.oliferation was MS medium supplemented with 0.5 mg!L IBA. The
growth of the roots increased from the first to the fourth subculture but decreased on
the fifth and sixth subculture. The in vitro roots of H. niger could also be established in . -
G).
150 mL and the one litre aeration tubes filled with 50 mL and 500 mL MS medium
supplemented with 0.5 mg/L IBA respectively. GC-MS analysis revealed the presence
of both hyoscyamine and scopolamine in all the H. niger roots which included roots of
the whole plant, roots induced on solid medium supplemented with 2.0 mg/L IBA,
roots produced on liquid medium supplemented with 0.5 mg!L ffiA on shaken flasks,
and roots produced in 150 mL and the one litre aeration tubes using MS medium
supplemented with 0.5 mg!L IDA Both hyoscyamine and scopolamine were not
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detected in the whole plant (with the roots separated), the, callus indu-ced on MS
medium supplemented with 6.0 mg!L of picloram, the liquid culture medium (MS + 0.5
mg!L IDA) from the shake flask and the liquid culture medium (MS + 0.5 mg!L IDA)
from the 150 mL aerated tube.
Description
Keywords
Root culture , Hyoscyanzus niger , Temperate medicinal plant